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The pH of the blood must be maintained within a very narrow range

For immunoprecipitation under denaturing conditions, cells were lysed in a urea-containing lysis buffer [10 mM Tris-HCl (pH, 7.5), 150 mM NaCl, 0.5 mM EDTA, 1% NP-40, 10% glycerol, and 6 M urea] supplemented with 20 M MG132, 10 M PR619, 10 mM iodoacetamide, and a protease inhibitor cocktail, and then extensively sonicated using Handy Sonic

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